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<div class="pagetitle">Cellular Response to Infection</div>

<div class="text">
    The in vivo infection studies are complemented by parallel work using cultures of primary airway epithelial cells,
    the predominant target cell of influenza. We have developed a three-dimensional mouse tracheal epithelial cell
    culture system (mTECs) that recapitulates much of the in vivo biology, including culture at the liquid-air
    interface, apical-basolateral polarization, tight junction formation, and beating cilia. Using this model system, we
    are determining the differences in the regulatory networks activated in the respiratory epithelium by strains of
    varying pathogenicity.
    <br/><br/>
    <a href="content/3D_airway_epithelial_culture.doc" target="_blank">3D Airway Epithelial Culture Protocol</a>
    <br/><br/>
    We have analyzed the response of mTECs to both high- and low-multiplicities of infection. The low multiplicity of
    infection (0.01) was chosen to match the number of cells initially infected in mice inoculated with 105 PFU of virus
    and primarily reveals the cellular response to a spreading infection. In the high MOI experiments (MOI = 2), the
    infection is synchronized across the entire cell population and the predominant transcriptional signature arises
    from infected cells.

    <div class="diagram">
        <img src="content/mTEC.png" alt="">

        <div class="legend">mTEC culture illustrating polarized monolayer growing on tissue culture plate insert
            suspended in media.
        </div>
    </div>
</div>

<div class="text">
    We are using <b>primary human airway epithelial (hAE) cells</b> (<a
        href="http://www.mattek.com/pages/products/epiairway">EpiAirway</a>&trade;, MatTek) to extend our studies to
    include
    current influenza strains circulating in the general population, most notably the recently emergent 2009
    Swine-origin Influenza (H1N1) virus (S-OIV). The pandemic strain appears to have arisen from a reassortant event
    between two circulating influenza strains that are endemic to swine in North America and Eurasia. While the two
    parent strains are not known to be human-transmissible, the particular combination of genes in the reassortant has
    produced a strain that has rapidly spread worldwide. In collaboration with the Center of Excellence for Influenza
    Research and Surveillance (CEIRS) at St. Jude Children's Research Hospital, we have initiated a systems-level
    analysis of the response of hAE cells to infection with this virus in comparison to the parent strains and to a
    seasonal influenza strain. The goal of this project is to understand the mechanisms by which the constellation of
    genes in the pandemic strain confers pathogenicity in humans in contrast with the parent strains. Additionally, we
    aim to uncover novel mechanisms of pathogenicity in the pandemic strain that are not present in circulating seasonal
    strains.
    <div class="diagram">
        <img src="content/hAEs.png" alt="">

        <div class="legend">Polarized hAE cells growing in culture</div>
    </div>
</div>

<div class="text">
    <h3>Microarray Data</h3>
    <br/>
    <h4>Affymetrix GeneChip Mouse Exon 1.0 ST Arrays of mTEC Cultures</h4>

    <div class="subtitle">
        Mouse tracheal epithelial cells were infected with the indicated strains at various multiplicities of infection
        (MOI) and RNA was extracted using TRIzol at the indicated time point. Samples were then amplified and hybridized
        to Affymetrix GeneChip Mouse Exon 1.0 ST Arrays.
    </div>

    <div id="c_mTEC"></div>

    <br/>
    <h4>Affymetrix GeneChip Human Exon 1.0 ST Arrays of hAE Cultures</h4>

    <div class="subtitle">
        Human airway epithelial cell cultures (<a href="http://www.mattek.com/pages/products/epiairway">EpiAirway</a>&trade;,
        MatTek) were infected with the indicated strains at MOI~0.01. RNA was extracted using TRIzol at the indicated
        time point and hybridized to Affymetrix GeneChip Human Exon 1.0 ST Arrays.
    </div>
    <div id="c_hAE"></div>
</div>

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